Oroxin A in Linggui Zhugan Decoction in the treatment of MASH by regulating cholesterol-bile acid metabolism.

ETHNOPHARMACOLOGICAL RELEVANCE: Lingui Zhugan Decoction (LGZGD), a traditional Chinese medicinal formulation documented in the Golden Cabinet Essentials, is widely employed across East Asia for the treatment of metabolic liver disease. AIM OF THE STUDY: To evaluate the anti - MASH efficacy of LGZGD and its Active blood-entering components and elucidate their mechanisms, with a particular focus on the regulation of bile acid metabolism. MATERIALS AND METHODS: A mouse model of MASH was established using a high-fat, high-cholesterol diet (HFHCD). Transcriptomic profiling and blood component analysis were performed on mice after LGZGD intervention. Molecular docking was then employed in combination with these analyses to screen for active constituents. Subsequent analyses focused on examining the effects of Oroxin A on MASH through transcriptomics, bile acid-targeted metabolomics, gut microbiota 16S rRNA sequencing, and Western blotting. RESULTS: LGZGD, Oroxin A all alleviated obesity, hepatic steatosis, inflammatory responses, and liver fibrosis induced by a high-fat diet. Microbial analysis of the ileum via 16S rDNA sequencing revealed that Oroxin A intervention restored gut microbial diversity, increasing the abundance of Paracoccus, Akkermansia, and Bifidobacterium. Targeted metabolomic analysis revealed significant alterations in bile acid composition under Oroxin A intervention, characterised by decreased levels of LCA and other bile acids alongside marked increases in TUDCA and others. Western blotting results indicate that Oroxin A applies its anti-MASH impacts by modulating the FXR/CYP7A1 pathway and TGR5/NLRP3 pathway. CONCLUSION: Oroxin A can improve pathological conditions in MASH mice through multiple pathways, including restoring gut microbiota balance and regulating bile acid metabolism. Within conventional medical frameworks, Oroxin A pioneers novel therapeutic approaches for MASH. This method overcomes the limitations of synthetic drugs while maintaining metabolic homeostasis.